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Thesis (Ph. D.)--University of Missouri--Columbia, 1998. / "July 1998." Typescript. Vita. Includes bibliographical references (leaves 145-165). Also available on the Internet.

Thesis (Ph. D.)--University of Missouri--Columbia, 1998. / Typescript. Vita. Includes bibliographical references (leaves: 145-165). Also available on the Internet.

Thesis (Ph. D.)--University of Missouri--Columbia, 1998. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (Ph. D.)--University of Missouri--Columbia, 1998. / Includes bibliographical references.

Chau Shuk Ling. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 103-115). / Abstracts in English and Chinese. / DECLARATION --- p.i / ACKNOWLEDGEMENT --- p.ii / ABBREVIATIONS --- p.iii / ABSTRACT IN ENGLISH --- p.v / ABSTRACT IN CHINESE --- p.viii / TABLE OF CONTENTS --- p.xi / Chapter CHAPTER I - --- INTRODUCTION / Chapter 1.1 --- Fluid Transport in Human Colon --- p.1 / Chapter 1.2 --- C1' Secretion across the Colonic Epithelium --- p.2 / Chapter 1.3 --- Properties of T84 Cells --- p.7 / Chapter 1.4 --- General Introduction on Extracellular Calcium Sensing Receptor (CaSR) --- p.8 / Chapter 1.5 --- Molecular Structure of CaSR --- p.9 / Chapter 1.6 --- CaSR-mediated Intracellular Signaling --- p.13 / Chapter 1.7 --- Agonists of CaSR --- p.16 / Chapter 1.8 --- Functions of CaSR --- p.18 / Chapter 1.8.1 --- Homeostatic Functions of CaSR --- p.18 / Chapter 1.8.2 --- Non-Homeostatic Functions of CaSR --- p.18 / Chapter 1.9 --- Molecular and Functional Study of CaSR Expressed in Colon --- p.19 / Chapter 1.10 --- Objectives of the Present Study --- p.21 / Chapter CHAPTER II - --- MATERIALS AND METHODS / Chapter 2.1 --- Solutions and Drugs --- p.22 / Chapter 2.2 --- Cell Culture --- p.23 / Chapter 2.3 --- Western Blot --- p.26 / Chapter 2.4 --- Simultaneous Measurement of Short-Circuit Current (Isc) and Intracellular Calcium Concentration ([Ca2+]i) --- p.27 / Chapter 2.4.1 --- Measurement of Isc and Transepithelial Resistance with Ussing Chamber --- p.27 / Chapter 2.4.2 --- Simultaneous Measurement of Isc and [Ca2+]i --- p.29 / Chapter 2.5 --- Measurement of Isc with Conventional Ussing Chamber --- p.32 / Chapter 2.6 --- Measurement of Intracellular cAMP Accumulation with Enzyme-Linked Immunosorbent Assay --- p.34 / Chapter 2.7 --- Data Analysis --- p.34 / Chapter CHAPTER III - --- RESULTS / Chapter 3.1 --- Expression of CaSR in T84 Cell Monolayers --- p.36 / Chapter 3.2 --- Poly L-arginine Induced an Increase in Isc --- p.38 / Chapter 3.2.1 --- Simultaneous Measurement of Isc and [Ca2+ ]i in Response to Poly L-arginine --- p.38 / Chapter 3.2.2 --- Interaction Between Poly L-arginine and Forskolin --- p.50 / Chapter 3.2.3 --- Ionic Mechanism of Isc Stimulated by Poly L-arginine --- p.58 / Chapter 3.2.3.1 --- Involvement of C1- in Isc Stimulated by Poly L-arginine --- p.58 / Chapter 3.2.3.2 --- Involvement of Basolateral K+ Channels in Isc Stimulated by Poly L-arginine --- p.61 / Chapter 3.2.4 --- Signaling Pathways Underlying the Isc Response to Poly L-arginine --- p.73 / Chapter CHAPTER IV - --- DISSCUSION / Chapter 4.1 --- Presence of CaSR in T84 Cell Monolayers --- p.78 / Chapter 4.2 --- Simultaneous Measurement of Isc and [Ca2+ ]i upon Application of Poly L-arginine --- p.82 / Chapter 4.3 --- Ionic Mechanism Underlying the Increase in Isc Stimulated by Poly L-arginine --- p.87 / Chapter 4.4 --- Signaling Pathway Underlying the Action of Poly L-arginine --- p.93 / Chapter 4.5 --- Does CaSR Mediate Poly L-arginine-evoked C1- Secretion across T84 Cell Monolayers? --- p.96 / Chapter 4.6 --- Future Study --- p.101 / REFERENCES --- p.103

[Truncated abstract] The calcium-sensing receptor (CaR) is a G protein-coupled receptor that can respond to changes in extracellular calcium and plays an integral role in calcium homeostasis. Later studies revealed that the CaR was stimulated by not just calcium, but a diverse range of stimuli and that activation of the receptor regulated a host of different biological processes. The CaR is linked to these cellular responses via the various signalling pathways initiated by the receptor. Recent yeast two-hybrid studies have identified a number of accessory proteins that, through their interaction with the intracellular tail of the CaR, are able to regulate important functional aspects of the receptor, including its signalling and degradation. We hypothesised that many more proteins that bind to the CaR-tail await identification, especially since most of the previous studies used the yeast two-hybrid system to screen cDNA libraries generated from tissues that are important to whole body calcium homeostasis, such as the parathyroid gland and kidney. In order to identify novel binding partners of the CaR, which may affect its function, particularly in biological processes that might be unrelated to calcium homeostasis, our laboratory performed a yeast two-hybrid screen of an EMLC.1 mouse pluripotent haemopoietic cell line library using the intracellular tail of the human CaR as bait. This screen revealed a large number of